In vitro cultivation of hairy roots of Plumbago rosea L. in a customized Reaction kettle for the production of plumbagin—An anticancer compound

•Established hairy root cultures of Plumbago rosea L.•A low cost reaction Kettle vessel (2 L) was customized for scaling up of hairy roots.•Hairy root biomass was increased twelve fold in 25 days.•Plumbagin content of hairy roots was enhanced to 1.5% DW in 25 days.

Biosynthesis of plumbagin using hairy root cultures of Plumbago rosea L. can be made economical only by scaling-up cultures in suitable vessels under conditions optimal for maximum yield of biomass and plumbagin. However, the major limitation faced in the scaling-up of hairy roots is the development of appropriate culture vessels that will allow maximum growth of interconnected root mat with less shear stress. Design of economical and low energy requiring scaling-up culture system for hairy roots with emphasis on physical parameters like inoculum state, nutrient mixing, aeration, inoculum support etc., is very important for root cultures. A novel, culture vessel model using a Reaction kettle (2 L) was attempted for successful scaling-up of P. rosea hairy roots. The roots cultured in Murashige and Skoog basal liquid medium for 25 days in a Reaction kettle (2 L) customized with a polypropylene mesh for support and a fish tank aerator for aeration yielded maximum biomass and plumbagin. Twelve-fold increase in root biomass with highest plumbagin content of 1.5% dry weight (DW) was obtained in Reaction kettle whereas, shake flask cultures produced only five fold biomass enhancement with 1.1% DW plumbagin. The present experiments demonstrated the suitability of the customized Reaction kettle with adequate filtered air for the scaling-up of hairy root cultures for secondary metabolite production. This is the first report on exploiting a Reaction kettle for in vitro scaling-up of organ cultures.

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